Cell cytotoxicity / Proliferation assay

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.

A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

Get tips on using QCM™ Tumor Cell Transendothelial Migration Assay (Colorimetric, 24 Assays) to perform Cell migration / Invasion cell type - SW480

Get tips on using CytoSelect™ 24-Well Cell Migration Assay, 8 μm to perform Cell migration / Invasion cell type - BCPAP

Get tips on using CytoSelect™ 24-Well Cell Migration Assay, 8 μm to perform Cell migration / Invasion cell type - SW480

Get tips on using CytoSelect™ 24-Well Cell Migration Assay, 8 μm to perform Cell migration / Invasion cell type - HUVEC

Get tips on using EZViable™ Calcein AM Cell Viability Assay Kit (Fluorometric) to perform Live / Dead assay mammalian cells - rat brain microvascular endothelial cells

Get tips on using Cultrex 3D Culture BME Cell Invasion Assay, 96 well to perform Cell migration / Invasion cell type - BxPC-3